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A mutation in the E2 subunit of the mitochondrial pyruvate dehydrogenase complex in Arabidopsis reduces plant organ size and enhances the accumulation of amino acids and intermediate products of the TCA Cycle

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Abstract

The mitochondrial pyruvate dehydrogenase complex (mtPDC) plays a pivotal role in controlling the entry of carbon into the tricarboxylic acid (TCA) cycle for energy production. This multi-enzyme complex consists of three components: E1, E2, and E3. In Arabidopsis, there are three genes, mtE2-1, mtE2-2, and mtE2-3, which encode the putative mtPDC E2 subunit but how each of them contributes to the total mtPDC activity remains unknown. In this work, we characterized an Arabidopsis mutant, m132, that has abnormal small organs. Molecular cloning indicated that the phenotype of m132 is caused by a mutation in the mtE2-1 gene, which results in a truncation of 109 amino acids at the C-terminus of the encoded protein. In m132, mtPDC activity is only 30% of the WT and ATP production is severely impaired. The mutation in the mtE2-1 gene also leads to the over-accumulation of most intermediate products of the TCA cycle and of all the amino acids for protein synthesis. Our results suggest that, among the three mtE2 genes, mtE2-1 is a major contributor to the function of Arabidopsis mtPDC and that the functional disruption of mtE2-1 profoundly affects plant growth and development, as well as its metabolism.

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Abbreviations

CAPS:

Cleaved amplified polymorphic sequences

DAG:

Days after germination

E3BP:

E3-Binding protein

iP:

N6-Isopentenyladenine

mtPDC:

Mitochondrial PDC

MS:

Murashige & Skoog

PDC:

Pyruvate dehydrogenase complex

qRT-PCR:

Quantitative real-time PCR

SSLP:

Simple sequence length polymorphism

TCA:

Tricarboxylic acid

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Acknowledgments

We thank Dr. Lijia Qu (Peking University) for providing Arabidopsis T-DNA activation library and Dr. Jianru Zhou (Institute of Genetics and Developmental Biology, Chinese Academy of Sciences) for insightful discussions about this manuscript. We are grateful to Mr. Yu Tian (Tsinghua University) for assistance on analysis of pyruvate content. This work was supported by the Ministry of Science and Technology of China (Grant no. 2009CB119100 to D. Liu), the National Natural Science Foundation of China (Grant no. 30971554 to X. Du), and the Ministry of Agriculture of China (Grant no. 2009ZX08009-123B to D. Liu).

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Authors and Affiliations

  1. MOE Key Laboratory of Bioinformatics, School of Life Sciences, Tsinghua University, Beijing, 100084, China

    Hailan Yu, Xiaoqiu Du, Fang Zhang & Dong Liu

  2. State Key Laboratory of Plant Genomics and National Plant Gene Research Center, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, 100101, China

    Fengxia Zhang & Guodong Wang

  3. College of Life Sciences, Capital Normal University, Beijing, 100048, China

    Yong Hu

  4. The Tree and Ornamental Plant Breeding and Biotechnology Laboratory of Chinese Forestry Administration, National Engineering Laboratory for Tree Breeding, College of Life Sciences and Biotechnology, Beijing Forestry University, Beijing, 100083, China

    Shichang Liu & Xiangning Jiang

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  1. Hailan Yu
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  2. Xiaoqiu Du
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  3. Fengxia Zhang
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  4. Fang Zhang
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  9. Dong Liu
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Corresponding author

Correspondence to Dong Liu.

Additional information

H. Yu, and X. Du contributed equally to this work.

The nucleotide sequence of mtE2-1 gene can be found in the TAIR database under the accession number At3g52200.

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Yu, H., Du, X., Zhang, F. et al. A mutation in the E2 subunit of the mitochondrial pyruvate dehydrogenase complex in Arabidopsis reduces plant organ size and enhances the accumulation of amino acids and intermediate products of the TCA Cycle. Planta 236, 387–399 (2012). https://doi.org/10.1007/s00425-012-1620-3

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